缺氧诱导因子-1在人乳腺癌细胞系MCF-7中的抗凋亡作用(英文)  

Anti-apoptotic role of HIF-1 in MCF-7 breast cancer cell line

作　　者：王晗[1] 符史干[1] 董战玲[1] 许闽广[1] 王杨[1] 陈国斌[1] 高凌峰[1] 

机构地区：[1]海南医学院生理学教研室,海南海口571101

出　　处：《中国热带医学》2011年第4期397-400,共4页China Tropical Medicine

基　　金：海南省自然科学基金(309035);海南省教育厅高校科研基金(Hjkj2009-55)

摘　　要：目的研究缺氧诱导因子-1在人乳腺癌细胞系MCF-7细胞中对凋亡的作用。方法氯化钴(CoCl2)模拟低氧环境。采用RNA干扰技术,构建针对HIF-1α的shRNA真核表达载体,转染MCF-7细胞。Real-time PCR和Western-blot技术分别检测HIF-1αmRNA和蛋白质水平。Sub-G1测定,AnnexinⅤ荧光标记和DNA ladder检测细胞凋亡。结果缺氧后MCF-7细胞HIF-1α的蛋白质水平表达增加,但缺氧后细胞的凋亡率比正常氧水平时低。实时定量PCR和Western blot结果证实转染短发夹RNA后,MCF-7细胞中HIF-1α基因表达被成功抑制。阿糖胞苷诱导或无凋亡诱导剂时,干扰组凋亡率比未转染组明显增高。结论这项研究证明HIF-1在MCF-7中发挥抗细胞凋亡的作用,为针对HIF-1α的shRNA有效地用于乳腺癌的治疗提供新的思路。HIF-1 （hypoxia-inducible factor-1 ） is the major transcriptional factor responsible for oxygen-dependent gene expression . Suppression of HIF-lot is important for exploring HIF-l-dependent pathophysiological processes. In this study, we investigated the effect of short hairpin RNA （shRNA）targeting HIF-lot on the human breast carcinoma MCF-7 cell line. The protein level of HIF-lot increased after exposure of MCF-7 cells to hypoxia. However,apoptosis was lower in hypoxia compared with normoxia. The results of real-time PCR and western blot validated that the HIF-lotgene of MCF-7 cells transfected with HIF-lot shRNA was suppressed successfully. When the cells were treated with or without apoptosis inducer Ara-c ,the apoptosis rate of MCF-7 cells transfected with HIF-lot shRNA was significantly higher than that of untransfected cells. Then the study proves the anti-apoptotic role of HIF-1 in MCF-7 breast cancer cell line and brings new insight into the strategy of using shRNA targeting HIF-letto treat breast cancer.

关 键 词：缺氧诱导因子-1Α 短发夹RNA 人乳腺癌细胞系 细胞凋亡 

分 类 号：R737.9[医药卫生—肿瘤]